Aggregate Removal by Responsive Electrospun Membrane based Hydrophobic Interaction Chromatography

Salt responsive polymer, poly(N-vinylcaprolactam), becomes hydrophobic in high salt concentration buffer. Electrospun cellulose nanofiber membranes grafted with PVCL polymer chains can separate protein monomer from its dimer as dimer has stronger hydrophobic interaction with PVCL chains than monomer.

Abstract

Poly(N-vinylcaprolactam) (PVCL), a salt responsive ligand, was grafted from electrospun cellulose nanofiber membranes by atom transfer radical polymerization (ATRP). The membranes were used for hydrophobic interaction chromatography. The modified PVCL-cellulose membranes were used to separate BSA dimers from monomers as well as monoclonal antibody (mAb) aggregates. Fast protein liquid chromatography (FPLC) was used for protein separation and fraction collection. High performance liquid chromatography with a size exclusion column (HPLC-SEC) analysis confirmed the separation of monomers and aggregates from both BSA and the mAb. The results suggest that use of a responsive ligand that changes its conformation with ionic strength could improve the performance of hydrophobic interaction chromatography enabling the separation of compounds based on slight differences in hydrophobicity.

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